Team:Paris Saclay/Notebook/July/30
Contents
Thursday 30th July
Lab Work
Transformation
by Audrey
Biobricks:
- BBa_K1707004
- BBa_K1707009
As usual
Plasmid extraction
by Johan
Biobricks:
- BBa_K1707008 #3, #4, #5 and #6
With the Macherey-Nagel Extraction kit
Ligation
by Pauline
- BBa_K1707011: BBa_B0030 + BBa_K1707010 #1
- 3 µL BBa_B0030
- 5 µL BBa_K1707010
- 2 µL Buffer Ligase 10x
- 1 µL Ligase
- BBa_K1707021: BBa_K1707000 + BBa_K1707010
- 1,5µL BBa_K1707000
- 5 µL BBa_K1707006
- 1 µL Buffer Ligase 10x
- 1 µL Ligase
- 1,5 µL H2O
Incubation 4h, 4°C
Digestion Verification
by Pauline
- Biobricks:
- BBa_K1707008 #2, #3, #4, #5, #6
- BBa_K1707005 #1 and #2
- Mix:
- 1 µL Buffer FastDigest 10x
- 0,5 µL XbaI
- 2 µL Plasmid
- 6,5 µL H2O
Incubation 1h30, 37°C
Electrophoresis
by Pauline
Agarose gel 1%, migration 80V then 110V
Verification by digestion with XbaI, from left to right: 1. DNA Ladder, 2. BBa_J23101, 3. BBa_K1707008#2, 4. BBa_K1707008#3, 5. BBa_K1707008#4, 6. BBa_K1707008#5, 7. BBa_K1707008#6, 8. BBa_K1707001#1, 9. BBa_K1707005#1, 10. BBa_K1707005#2, 11. Empty, 12. Empty
We can conclude that BBa_K1707008 isn't OK, and BBa_K1707005 #1 and #2 are OKNew Culture
by Pauline
Biobricks:
- BBa_K1707012
- BBa_K1707013
- BBa_K1707019
4 clones for each, in 5ML LB + 20 ng/µL Chloramphenicol
Transformation
by Johan and Pauline
Biobricks:
- BBa_K1707021
- BBa_K1707011
- BBa_I13600
As usual
Ligation
by Coralie
- BBa_K1707008: BBa_J23101 + BBa_K1707003
- 2,5 µL BBa_J23101
- 11 µL BBa_K1707003
- 2 µL Buffer Ligase 10x
- 1 µL Ligase
- 3,5 µL H2O
Incubation over night, 16°C
Soil experiment
by Coralie
Soil
Observation of J0 plates: there are a lot of colony in each. Contamination is alright. Controls (+) and (-) are Ok too. We take 1g of each pot of soil and we dilute it in 5mL sterile H2O. After shaking and decant, we put 100µL of supernatant on the right plate. Incubation ON 37°C
Interlab study: Cytometer
by Johan
https://2015.igem.org/Team:Paris_Saclay/Measurement#30th_July
Member present:
- Instructors: Alice
- Students: Coralie, Audrey, Pauline, Johan and Seong-Koo